Lipid conjugates of therapeutic peptides and protease inhibitors

ABSTRACT

Compounds wherein therapeutic peptides are covalently linked to phospholipids, glycerides or other membrane-targeting and membrane-anchoring species, and their pharmaceutically acceptable salts, together with processes for their preparation. The targeting technology is applicable to HIV protease inhibitors and a variety of other enzyme inhibitors.

This invention provides lipid conjugates of therapeutic peptides. Italso provides peptide inhibitors of HIV protease.

BACKGROUND OF THE INVENTION

The uptake of therapeutic peptides into cells following in vivoadministration is not efficient. A substantial amount of peptides areremoved extremely rapidly from the plasma by a variety of mechanismsincluding cellular uptake and metabolism, filtration by the kidney andurinary excretion or destruction by the proximal tubule cells of thekidney. Furthermore the lipid bilayer of the cell membrane presents abarrier to transport. Yet another disadvantage of these peptides is theneed to administer them by injection because peptides in general possessvery poor oral or nasal bioavailability.

The human immunodeficiency virus (HIV) produces a long gag polyproteinwhich is cleaved during viral budding into smaller proteins which havespecific viral functions. This proteolytic cleavage is catalyzed by aspecific HIV aspartyl protease enzyme (HIV PR). Small peptide fragmentswhich conformationally resemble the HIV protease binding site but havethe p1 site replaced by non-hydrolyzable structures act as inhibitors ofthe HIV protease and may be useful as therapeutic agents in AIDS.

A number of such protease inhibitors have been synthesized. See forexample, Merck European Patent Application # EP 0 337 714 A,publications by Upjohn, Science 247:454-456 (1990), Roche EuropeanPatent Application EP 0 346 847 A2, Smith, Kline French InternationalPatent Application WO 90/00399, Nature 343:90-92 (1990), Proc. Natl.Acad Sci.USA 86:9752-9756 (1989), Hoechst European patent Application EP0 354 522 A1. All of these protease-inhibitors are peptides, and aspeptides, are subject to a general problem of metabolic degradation andclearance which will prevent them from reaching the infected celltargets to bind HIV PR.

Protease inhibitory peptides, as well as other therapeutic peptides,lack targeting mechanisms in their native form, and on administration toan animal they become distributed globally. For that reason they may noteffectively treat infected tissues. For example, macrophages arebelieved to be a major site of HIV infection, and inhibitory peptides bythemselves do not target to the macrophage reservoir.

These problems can be overcome by preparing conjugates of therapeuticpeptides which resist clearance and degradation, and which can betargeted to the cell.

It is therefore the object of the invention to overcome the problem ofdegradation and clearance of therapeutic peptides, including HIVprotease inhibitory peptides, from the plasma. It is further the objectof the invention to provide peptides comprising nonhydrolyzable groupscapable of inhibiting HIV PR. Another object of this invention is toprovide for prodrugs of protease inhibitory peptides which treat thesite of viral infection effectively. Yet another object of thisinvention is to improve the oral and nasal bioavailability oftherapeutic peptides.

SUMMARY OF THE INVENTION

The invention provides peptide-lipid linking structures, proteaseinhibitors, and compounds wherein protease inhibitors and othertherapeutic peptides are covalently linked to phospholipids, glyceridesor other cell membrane targeting or membrane anchoring moieties.

According to one aspect of the invention there are provided therapeuticpeptides which are analogues of substrates for proteases. These peptidesinclude the protease substrates of Group I described in detail in thespecification. Preferred peptides of this group inhibit the HIVprotease.

According to another aspect of the invention there are providedcompounds capable of linking peptides to lipid structures. These linkingcompounds include the amino acid phospholipids of Group IIA-D describedin the specification, as well as other lipid-bearing linkers defined asX species, suitable for attachment to an amino group of a peptide and Cspecies, suitable for attachment to the carboxyl group of a peptide.

According to yet another aspect of the invention there are providedpeptide-lipid conjugates, comprising therapeutic peptides conjugated toany of the lipid-bearing linkers defined by the invention. Preferredspecies are lipid derivatives of protease inhibitory peptides. Thepeptide lipid conjugates can also comprise spacers, comprisingbifunctional alkyl groups defined as Y, Z, and W species, which canmodulate the distance between the peptide and the lipid linker.Preferred peptide-lipid conjugates comprise peptides that are proteaseinhibitors. These conjugates of Group III-A-F are described in detail inthe specification.

The invention also provides intermediate compounds useful in thesynthesis of the peptide-lipid compounds of the invention, comprisingderivatized linking compounds suitable for binding to the functionalgroups of lipids or peptides.

The invention also provides a method for synthesizing peptide-lipidconjugates, comprising the steps of selecting a therapeutic peptidehaving an available amino group or carboxyl group; chemically bonding abivalent linker group to either the amino or the carboxyl group; andchemically bonding a lipid species to the linker. In a preferred method,the peptide is a protease inhibitor.

The invention further provides a method of treating infectious diseasecaused by a virus, comprising administering to the infected person aneffective virus-inhibiting amount of a lipid derivative of a viralprotease inhibitory peptide. The inhibitory peptide-lipid can beincorporated into a liposome prior to administration. In a preferredembodiment of the method, the disease treated is HIV infection, and thepeptide is a HIV protease inhibitor.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides protease inhibitory peptides, lipid conjugates oftherapeutic peptides, particularly peptides that are enzyme substrates,such as the HIV protease inhibitory peptides, linkers capable ofattaching lipid species to the peptides, intermediate compounds, andmethods of synthesis and use.

Group I: Protease inhibitors:

Protease inhibitors (PIs) are, in general, substrates of HIV 1 proteasein which the amino acid residues at the p1 site are replaced byisosteric residues. Some of the HIV protease substrates which are thebasis for the design of PIs are:

    ______________________________________                                        Sequence              Site                                                    ______________________________________                                        Ser--Gln--Asn--Tyr--Pro--Ile--Val                                                                   p17/p24                                                 Ala--Arg--Val--Leu--Ala--Glu--Ala                                                                   p24/p7                                                  Ala--Thr--Ile--Met--Met--Gln--Arg                                                                   p24/p7                                                  Ser--Phe--Asn--Phe--Pro--Gln--Ile                                                                   HIV PR N-term                                           ______________________________________                                    

Detailed descriptions of the design and structures of individual PIs arediscussed below. The present invention provides for the following PIs:

Ser-Gln-Asn-Phe-Pro-Ile-Val-NH₂ ;

Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂ ;

Ser-Gln-Asn-Tyr-Achx-Ile-Val-NH₂ ;

Ser-Gln-Asn-Tyr-Acpr-Ile-Val-NH₂ ;

Ser-Gln-Asn-Tyr-Acpnt-Ile-Val-NH₂ ;

Thr-Ile-Leu-(beta-Ala)-Leu-Gln-Arg-NH₂ ;

Ser-Gln-Asn-Tyr-Pro-Ile-Val-Thr-Leu-Ava-Thr-Gln-Arg-NH₂

Ac-Ala-Ala-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂ ;

Ac-Ala-Ala-Phe-Pip-(a-(OH)-Leu)-Val-NH₂ ;

Ac-Ala-Ala-(DL-Phe(4-Cl))-Pip-(a-(OH)-Leu)-Val-NH₂ ;

Ala-Ala-Phe-(beta-Ala)-Val-Val-Gly-OH;

Ala-Ala-Phe-(beta-Ala)-Nva-Val-Gly-OH;

Ala-Ala-Phe-(beta-Ala)-(a-(OH)-iso-Valeryl)-Val-Gly-OH;

Ala-Ala-Phe-(beta-Ala)-Val-Val-Gly-OMe;

Ala-Ala-Phe-(beta-Ala)-Nva-Val-Gly-OMe;

Ala-Ala-Phe-(beta-Ala)-(a-(OH)-iso-Valeryl)-Val-Gly-OMe;

Boc-Ala-Ala-Phe-(beta-Ala)-Nva-Gly-OMe;

(a,e-di-Myristoyl-Lys)-D-Pro-Ava-Ala-Phe-Ava-Val-Gly-OMe

Ac-Ala-Ala-(D-a-Nal)-Pip-OMe;

Ac-Ala-Ala-(D-a-Nal)-Pip-NHNH₂ ;

Ac-Ala-Ala-(L-a-Nal)-Pip-OMe;

Ac-Ala-Ala-(L-a-Nal)-Pip-NHNH₂ ;

Ac-Ala-Ala-(D-b-Nal)-Pip-OMe;

Ac-Ala-Ala-(D-b-Nal)-Pip-NHNH₂ ;

Chl-Ala-Ala-(L-a-Nal)-Pip-OMe;

Chl-Ala-Ala-(L-a-Nal)-Pip-NHNH₂ ;

Paa-Ala-Ala-(D-a-Nal)-Pip-OMe;

Ac-Ala-Ala-(DL-Phe(4-Cl))-Pip-OH;

Val-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂ ;

Val-Ser-Gln-Asn-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂ ;

i-Boc-(D-Phe)-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂ ;

(i-Boc-(D-Phe)-Leu-)₂ -(3,5-di-Aba)-Pro-Leu-(D-Phe)-NH₂ ;

(Myristoyl-(D-Phe)-Leu-)₂ -(3,5-di-Aba)-Pro-Leu-(D-Phe)-NH₂ ;

i-Boc-(3,5-di-Aba)-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂ ;

Pro-Ile-Val-NH₂ ;

Asn-Phe(CO--CH₂ N)Pip-Ile-NH₂ ;

Quinaldoyl-Asn-Phe(CO--CH₂ N)Pip-Ile-NH₂ ;

Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂ ;

Quinaldoyl-Asn-Phe(CHOH--CH₂ N)Diq-Ile-Val-Gln-NH₂ ;

Val-Ser-Gln-Asn-Tyr-Diq-Ile-Val-Gln-NH₂ ;

Boc-Asn-Phe(CO--CH₂ N)-Diq-NHtBu;

Succinyl-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂ ;

Succinyl-Ser-Gln-Asn-Tyr;

Quinaldoyl-Asn-Phe(CHOH--CH₂ N)Pip-Ile-NH₂ ;

Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂ ;

Phe(CHOH--CH₂ N)Pro-Ile-Val-NH₂ ;

His-Lys-Arg-Ala-Val-Leu-Phe(4-NO₂)-Glu-Ala-Nle-Ser-NH₂ ;

[D-Phe]-[D-a-Nal]-Pip-[L-a-(OH)-Leu]-Val-NH₂ ;

Boc-Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂ ;

Boc-Phe(CHOH--CH₂ N)Pro-Ile-Val-NH₂ ;

Succinyl-Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂ ;

[D-Phe]-[D-b-Nal]-Pip-[L-a-(OH)-Leu]-Val-NH₂ ;

i-Boc-Tyr-Pro-Ile-Gly-OH;

(Boc-Phe-O-CH₂ --CHOH)₂ ;

(Boc-Val-Phe-O--CH₂ --CHOH)₂ ;

(Boc-Ser-Phe-O--CH₂ --CHOH)₂ ;

(Boc-Asn-Phe-O--CH₂ --CHOH)₂ ; and

(Boc-Arg-Phe-O--CH₂ --CHOH)₂.

Lipid bearing Linkers

The invention also provides a means for improving the usefulness,efficacy, biological half life, transport across cellular membrane, oraland or nasal bioavailabilities of therapeutic peptides, including viralprotease inhibitors by conjugating them to lipids via ester, amide,phosphate or phosphodiester bonds.

The protease inhibitory peptides of the invention, as well as othertherapeutic peptides, can be derivatized with groups which help toincrease their plasma half life and protect against tissue and renalclearance. By selecting suitable anchoring groups, that is membraneseeking, lipid associating groups, one can achieve selective targetingof the peptide or protease inhibitor. A number of natural and syntheticmolecules such as glycolipids, phospholipids, fatty acids, diglycerides,and lipid associating groups such as cholesterol, lend themselves ascandidates for anchoring the protease inhibitors to the lipid bilayer ofcell membranes.

Furthermore, the linkage between the anchoring group and the peptide orprotease inhibitor can be modulated in such a way as to obtain optimumrate of release of the drug or to position the drug in the most suitableconformation. These modified analogues of the viral protease inhibitorypeptides may then be incorporated into liposomes, administered byinjection (subcutaneously, intravenously, intraperitoneally orintramuscularly) to an animal. The derivatized peptides in liposomeswill then have a much longer time of retention in the plasma; and willsurvive to be delivered to macrophages, a major site of HIV infection.In addition it can be expected that the lymphocyte reservoir of the HIVinfection will also receive a larger amount of the lipid-modifiedpeptide by virtue of its ability to undergo exchange and transfer to thesurface membranes of cells in the circulation, such as the CD4+lymphocytes. Finally, this approach will also be usefully applied toother protease inhibitory peptides, such as those for renin.

In order to prepare peptide-lipid conjugates, a number of linker groupswere designed. They offer a convenient way of conjugating lipids,phospholipids, fatty acids, or glycerides to small peptides or proteaseinhibitors. These linkers are designed to take advantage of naturalbiochemical processes to release the active drug at or near the site ofenzyme action thus imparting improved biological functions to the parentdrug. A number of molecules containing multiple functional groups aresuitable these linkers.

Particularly suitable for use as linkers are the amino acids havinghydroxyl functional groups. These hydroxyamino acids are attached tophosphate or the phosphate group of phospholipids through a phosphoesterbond between the phosphate group and the amino acid hydroxyl, thusleaving both characteristic amino acid groups, the amino acid NH₂, andthe amino acid COOH, available to attach to the peptide carboxyl andamino groups, respectively, through amide bonds. Hydroxyamino acidphosphates can be attached to diacyl, dialkyl, monoacyl or monoalkylglycerols to provide peptides linked to lipids via a monophosphate bond.Phosphatidyltyrosine, phosphatidyl-serine, phosphatidyl-threonine, andphosphatidyl-hydroxyproline are some examples of preferred linkers. In asimilar manner diacyl, dialkyl, monoacyl or monoalkyl phospholipidscould also be attached to the phosphatidyl-hydroxyamino acids to furnishpeptides linked to lipids through a diphosphate bond.

The following linker moieties are preferred for preparing thepeptide-lipid conjugates:

Group II: Lipid-bearing Linker compounds

The invention provides phospholipid-amino acid compounds, having thephosphoester bond described above, wherein the phospholipid comprisesone or two linear or branched C₄ -C₂₄ alkyl chains, each chain havingfrom 0 to 6 double bonds. C₁₂ -C₁₈ alkyl chains are preferred, and thosehaving an even number of carbon atoms particularly preferred. The alkylchains can be attached by means of the hydroxyl groups of the glycerylportion of the phospholipid in 1,2-diradyl form, that is throughacyl/acyl acyl/alkyl alkyl/acyl or alkyl/alkyl bonds. Alternatively, thephospholipid may be a lyso species, having only one alkyl chain.

Accordingly, preferred species of lipid bearing linkers are:

Group II-A: 1,2-diradyl-sn-glycero-3-phospho-O-tyrosine,1,2-diradyl-sn-glycero-3-phospho-O-serine,1,2-diradyl-sn-glycero-3-phospho-O-hydroxyproline and1,2-diradyl-sn-glycero-3-phospho-O-threonine;

Group II-B: 1,2-diradyl-sn-glycero-3-diphospho-O-tyrosine,1,2-diradyl-sn-glycero-3-diphospho-O-serine,1,2-diradyl-sn-glycero-3-diphospho-O-hydroxyproline and1,2-diradyl-sn-glycero-3-diphospho-O-threonine;

Group II-C: 1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline and1-O-acyl-sn-glycero-3-phospho-O-threonine; and

Group II-D: 1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyprolineand 1-O-acyl-sn-glycero-3-diphospho-O-threonine.

Group II-E: 1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline, and1-O-alkyl-sn-glycero-3-phospho-O-threonine; and

Group II-F: 1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,and 1-O-alkyl-sn-glycero-3-diphospho-O-threonine.

Particularly preferred species of Group II-A are: phosphatidyl-tyrosine,phosphatidyl-serine, phosphatidylthreonine, andphosphatidyl-hydroxyproline.

Particularly preferred species of Group IIB are: tyrosine-O-diphosphatedipalmitoylglycerol, serine-O-diphosphate dipalmitoylglycerol,hydroxyproline-O-diphosphate dipalmitoylglycerol andthreonine-O-diphosphate dipalmitoylglycerol;

Particularly preferred species of Group II-E are:1-O-hexadecyl-sn-glycero-3-phospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-phospho-O-serine,1-O-hexadecyl-sn-glycero-3-phospho-O-hydroxyproline and1-O-hexadecyl-sn-glycero-3-phospho-O-threonine; and

Particularly preferred species of Group II-F are:1-O-hexadecyl-sn-glycero-3-diphospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-diphospho-O-serine,1-O-hexadecyl-sn-glycero-3-diphospho-O-hydroxyprolineand 1-O-hexadecyl-sn-glycero-3-diphospho-O-threonine.

Spacers

The distance between the lipid linker and the peptide can be adjusted bythe insertion of spacer moieties. Spacers suitable for use in thepeptide-lipids of the invention have two functional groups capable ofbinding to functional groups present on the lipid linkers and peptidesand serving to conjugate them.

Preferred spacers are:

W: H₂ N--(CH₂)_(n) --COOH, wherein n=1 to 12, suitable for insertion atthe carboxyl terminal of a peptide, connecting it with a lipid linkerand those moieties defined below as C.

Y: HOOC--(CH₂)_(n) --COOH, wherein n=1 to 12, suitable for insertion atthe amino terminal end of a peptide, to connect it with a phospholipidhaving an amino functional group. See, for example, in Group III-Abelow, wherein succinyl is a spacer connecting phosphatidyl-ethanolaminespecies DPPE and DMPE with N-terminal amino acids of the peptidespecies.

Z: HO--(CH₂)_(n) --COOH, wherein n=1 to 12, suitable for insertion atthe amino terminal end of a peptide, to connect the peptide with aphospholipid having a phosphate terminal group, for example,phosphatidic acid species such as DMPA and DPPA.

Group III: Peptide-lipid conjugates

In order to improve the biological lives of the inhibitors in vivo andto achieve their optimum targeting and release, we have designed thefollowing series of derivatives of protease inhibitory peptides. Anumber of linkages between phospholipids and peptides are possible. Forexample, phosphatidic acid can be linked to the amino or the hydroxylgroups of peptides. Alternatively, carboxyl groups of the peptides canbe attached to the amino group of a phosphatidyl-ethanolamine.

Yet another strategy for obtaining the peptide phospholipid conjugatesis to use suitable linkers, such as those comprising hydroxyamino acidsas described previously. A variety of other moieties lend themselves assuitable linkers for conjugating peptides to lipids.

The following peptide-lipid conjugates are described as examples, othertherapeutic peptides and enzyme inhibitors similarly can be convenientlyderivatized with appropriate lipid moieties by following the proceduresprovided in the present invention.

Group III-A:

The following are examples of peptide-lipid conjugates wherein thelipid, usually a phospholipid, or species of the lipid linkers describedabove (X), is attached at the terminal amino group of the peptide.##STR1##

Abbreviations:

Ac=Acetyl; Boc=t-Butyloxycarbonyl; Sucsuccinic acid; --OMe=Methyl ester,--NH₂ =Amide; --NHOH=Hydroxylamide; --NHNH₂ =Hydrazide;DPPA=1,2-di-Palmitoyl-phosphatidic acid;DPPE=1,2-di-Palmitoylphosphatidyl ethanolamine; Achx=1-Amino,1-cyclohexane carboxylic acid; Acpr=1-Amino, 1-cyclopropane carboxylicacid; Acpnt=1-Amino, 1-cyclopentane carboxylic acid; Pip=Pipecolic acid(4-piperidine carboxylic acid); Ava=5-Amino valeric acid,a-(OH)-Leu=L-Leucic acid (2-OH-L-isocaproic acid); Nal=Naphthylalanine;Phe(4-Cl)=p-Chloro phenylalanine; Nva=norvaline; Paa=Phosphonoaceticacid; Chl=Cholic acid. 3,5-di-Aba=3,5-di-Aminobenzoic acid.

X represents phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-hexadecyl-sn-glycero-3-phospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-phospho-O-serine,1-O-hexadecyl-sn-glycero-3-phospho-O-hydroxyproline,1-O-hexadecyl-sn-glycero-3-phospho-O-threonine,1-O-hexadecyl-sn-glycero-3-diphospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-diphospho-serine,1-O-hexadecyl-sn-glycero-3-diphosphohydroxyproline,1-O-hexadecyl-sn-glycero-3-diphosphothreonine, or DPPE-Y or DPPA-Z,

wherein Y is HOOC--(CH₂)_(n) --COOH;

Z is HO--(CH₂)_(n) --COOH; and n=1 to 12.

The linker moieties described here are useful for conjugating lipidgroups to a number of therapeutic peptides and protease inhibitorsdescribed in, for example, the following publications or patentapplications: U.S. Pat. No. 4,743,585; DE 3913272; DE 3840452; DE3819846; DE 3800233; EP 0 387 231; EP 0 337 714; EP 0 354 522; EP 0 356223; EP 0 373 576; EP 0 373 549; EP 0 372 537; EP 0 365 992; EP 0 361341; EP 0 386 611; EP 0 357 332; EP 0 342 541; EP 0 337 714; FR 2646353;WO 9101327; WO 9012804; WO 00399; WO 8910920, WO 8809815. The peptidesand the procedures described for the preparation of the said compoundsand related intermediates are incorporated herein by reference.

In the examples that follow, X and the abbreviations are as definedabove for Group III-A, and

C represents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-hexadecyl-sn-glycero-3-phospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-phosphoserine,1-O-hexadecyl-sn-glycero-3-phospho-O-hydroxyproline and1-O-hexadecyl-sn-glycero-3-phospho-O-threonine,1-O-hexadecyl-sn-glycero-3-diphospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-diphospho-O-serine,1-O-hexadecyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-hexadecyl-sn-glycero-3-diphospho-O-threonine, or DPPE-W, wherein Wis H₂ N--(CH₂)_(n) --COOH, and n=1 to 12.

Group III-B: Peptide-lipid conjugates based on the peptides of MerckEuropean Patent application # EP 0 337 714 A2

The following are examples of peptide-lipid conjugates wherein thelipid, a species of the lipid linkers described above (X), is attachedat the terminal amino group of the peptide or a species (C) is attachedat the carboxyl end of the peptide. In preferred species of this class,the peptide is linked to one lipid species, thus X is present only whenC is absent or is not a lipid. When C is present, it can be separatedfrom the peptide by the insertion of a W type spacer, described above.When X is present, it can be separated from the peptide by a Y or Z typespacer, also described above.

5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-Phe-C;

5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-C;

5(S)-X-amino-4(S)-hydroxy-6-cyclohexyl-2(R)-(phenylmethyl)hexanoyl-C;

5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-Phe-C;

5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-C;and

5(S)-X-amino-4(R)-hydroxy-6-cyclohexyl-2(R)-(phenylmethyl)hexanoyl-C.

wherein X and C are as defined above.

Group III-C: Peptide-lipid conjugates based on the peptides of UpjohnPublications: SCIENCE, VOL 247 PP 454-456, 1990; and Sawyer, T. K. etal. (1990) in Peptides: Chemistry Structure and Biology (Rivier, J. E. &Garland, G. R. eds.), pp. 855-857

The following are examples of peptide-lipid conjugates of the same typeas those presented in Group IIIB.

5(S)-X-Amino-4(S)-hydroxy-6-cyclcohexyl-2(R)-isopropyl-hexanoyl-Ile-C;

5(S)-X-Amino-4(R)-hydroxy-6-cyclcohexyl-2(S)-isopropyl-hexanoyl-Ile-C;

5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-isopropyl-hexanoyl-Ile-C;

5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(S)-isopropyl-hexanoyl-Ile-C;

wherein X and C are as defined above.

Group III-D: Peptide-lipid conjugates based on the peptides of the ROCHEEuropean Patent Application #EP 0 346 847 A2

The following are examples of peptide-lipid conjugates of the same typeas those presented in Group IIIB.

(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-Pro-C;

(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-Pro-C;

(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-C;

(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-Ile-C;

(3(S)-X-Leucyl-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-Ile-C;

(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;

(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;

(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;

(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C;

(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C; and

(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C;

wherein X and C are as defined above.

Group III-E: Peptide-lipid conjugates based on the peptides of the SK&FInternational Patent Application # WO 90/00399; and Publications: NATURE343:90-92 (1990); PNAS, VOL 86:9752-9756 (1989):

The following are examples of peptide-lipid conjugates of the same typeas those presented in Group III-B.

4(S)-(X-Alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

4(S)-(X-Alanyl-alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

4(S)-(X-Seryl-alanyl-alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

4(S)-(X-Alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

4(S)-(X-Alanyl-alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

4(S)-(X-Seryl-alanyl-alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C

wherein X and C are as defined above.

Group III-F: Peptide-lipid conjugates based on the peptides of theHOECHST European Patent # EP 0354 522 A1

The following are examples of peptide-lipid conjugates of the same typeas those presented in Group III-B.

X-Val-Phe-Nva-(cyclohexylmethyl(4,4,5,5-tetramethyl-1,3,2-dioxoborlan-2-yl)methylamide;

X-Val-Phe-Nva-(cyclohexylmethyl,dihydroxyboronyl)methylamide;

X-Val-(L-a-Nal)-Nva-(cyclohexylmethyl(4,4,5,5-tetramethyl-1,3,2-dioxoborlan-2-yl)methylamide;

X-Val-(L-a-Nal)-Nva-(cyclohexylmethyl,dihydroxyboronyl)methylamide;

X-Nva-(cyclohexylmethyl,dihydroxyboronyl)methylamide;

X-Val-(cyclohexylmethyl,dihydroxyboronyl)methylamide;

wherein X is as defined above.

As described in the experimental section, many of the lipid linkerspecies, particularly a phosphatidylhydroxyamino acid, are bifunctional,and can be positioned either at the C-terminal, N-terminal or in themiddle at internal sites of the protease inhibitory peptide as desired.That species of lipid linker can also be attached to two sites of thesame peptide, using appropriate spacers, if necessary. It is also withinthe contemplation of the invention to conjugate a monoglyceride ordiglyceride directly to a free carboxyl, preferably the terminalcarboxyl, of a peptide through the hydroxy group of the lipid.

Peptide Synthesis

Peptides of the invention can be produced by any of the peptidesynthesis procedures known to those in the art, for example, solutionphase synthesis, fragment condensation, enzyme synthesis, or any of themethods of solid phase synthesis. These peptides can also be produced byrecombinant DNA technology. The solution phase method and the solidphase methods are preferred. The solid phase methods are particularlypreferred. These methods are well known to those skilled in the art anddescribed in detail in the literature; e.g. Barany, G. and R. B.Merrifield, in The Peptides, vol. 2; E. Gross & J. Meienhoffer, eds.;Academic Press, New York, pp 3-284 (1979). Commercially availablederivatized amino acids used for the synthesis of peptides described inthis invention are: Boc-Ala-OH, Boc-Arg(Tos)-OH, Boc-Asn-OH,Boc-Asp(O-cyclohexyl)-OH, Boc-Asp(OBzl)-OH, Boc-Cys(S-4-MeBzl)-OH,Boc-Gln-OH, Boc-Glu(O-cyclohexyl)-OH, Boc-Glu(OBzl)-OH, Boc-Gly-OH,Boc-His(Tos)-OH, Boc-His(Bom)-OH, Boc-Ile-OH, Boc-Leu-OH,Boc-Lys(Cl-Z)-OH, Boc-Norleucine, Boc-Norvaline, Boc-Met-OH, Boc-Phe-OH,Boc-Pro-OH, Boc-Ser(Bzl)-OH, Boc-Thr(Bzl)-OH, Boc-Thr(Bzl)-OH,Boc-Trp-OH, Boc-Trp(Formyl)-OH, Boc-Tyr(Br-Z)-OH, and Boc-Val-OH.

Therapeutic applications

Peptides of the invention containing basic amino acids such as lysine,arginine, and histidine may exist in the form of salts such as chloride,acetate, phosphate, citrate, succinate, oxalate, etc. Acetate andhydrochloride salt forms are particularly preferred. Peptides of theinvention containing aspartic acid, glutamic acids or phosphate linkermoieties may exist in the form of salts such as sodium, potassium,calcium, barium, ammonium or other acceptable cation. For the purposesof this invention, peptides of the invention and their acid additionsalts are considered to be one and the same.

The peptides and peptide-lipid conjugates of the present invention areuseful as inhibitors of viral proteases and therefore could be used astherapeutic agents for example for the prevention or treatment ofinfection by human immunodeficiency virus (HIV), and subsequent diseaseconditions such as acquired immunodeficiency syndrome (AIDS) and AIDSrelated complex (ARC).

The peptide lipid conjugates are also useful in improving the efficacyof other protease inhibitors, such as inhibitors of renin. Renin is aprotease of approximately 40,000 MW secreted in the kidney byjuxtaglomerular cells surrounding the afferent arterioles of thecortical glomeruli. Renin itself has no activity, but acts on a proteinsubstrate, angiotensinogen, to split off the inactive decapeptide,angiotensin I, which is then converted through the splitting off of twoC-terminal peptides, to the active pressor agent, the octapeptideangiotensin-II. Angiotensin-II is the most potent pressor made in thebody and it exerts this pressor action by a direct effect on arteriolarsmooth muscle. Secretion of renin is stimulated in various disorders andcauses hypertension. An approach to the therapy of hypertensioncomprises the administration of renin inhibitory peptides which arerenin substrate analogues, a strategy comparable to the administrationof HIV protease inhibitors in the therapy of HIV infection. Lipidconjugates of renin inhibitory peptides, having the composition of thepeptide-lipids of the invention, can similarly contribute to theeffective therapy of hypertension by providing renin peptides in a formthat resists clearance and degradation and promotes efficient uptake bythe cells.

The present invention also relates to the use of the therapeuticpeptides and peptide conjugates of the invention and theirphysiologically acceptable salts for the preparation of pharmaceuticalformulations which can be employed as medicaments in human andveterinary medicine. For this purpose, it is possible to convert theminto a suitable form for administration together with at least onevehicle or auxiliary. Suitable vehicles are organic and inorganicsubstances which are suitable for enteral (for example, oral),parenteral, topical, transdermal or nasal administration and which donot react with the active drug substances. The indicated formulationscan be sterilized and/or contain auxiliaries, such as lubricants,preservatives stabilizers, wetting agents, emulsifiers, buffers,colorings and flavoring.

The active peptides may be administered parenterally, that is bysubcutaneous, intramuscular, intraperitoneal, or intravenous injection.The pharmaceutical formulations suitable for injectable use includeaqueous solutions or dispersions and powders for the extemporaneouspreparation of injectable solutions or dispersions. It is possible alsoto freeze-dry the peptides and to use the lyophilizates obtained, forexample, for the preparation of products for injection. In all cases,the form must be sterile and the solution must be fluid to the extentthat easy syringability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against contaminationsof microorganisms, such as for example, bacteria and fungi. The carriercan be a solvent or a dispersion medium containing, for example, water,or a polyol such as glycerol, and suitable mixtures thereof.Compositions for intramuscular use may also contain minor amounts ofsalts, acids, and bases to adjust tonicity and buffer the solution.Suitable buffering and isotonicity agents are readily determinable bypersons skilled in the art.

Oral or nasal administration is also possible especially withpeptide-lipid conjugates. Formulations for oral ingestion are in theform of tablets, capsules, pills, ampoules of powdered active agent, oroily or aqueous suspensions or solutions. Tablets or other non-liquidoral compositions may contain acceptable excipients, known to the artfor the manufacture of pharmaceutical compositions, comprising diluents,such as lactose or calcium carbonate; binding agents such as gelatin orstarch; and one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring or preserving agents toprovide a palatable preparation. Moreover, such oral preparations may becoated by known techniques to further delay disintegration andabsorption in the intestinal tract. Such oral compositions andpreparations should contain at least 0.1% of active peptide, althoughthe percentages of the compositions may vary widely. The amount oftherapeutically active compound in such compositions is such that asuitable dosage will be obtained in a convenient volume for ingestion.

Formulations for nasal administration may be in the form of liquids andoptionally may contain absorption promoting substances, for example, alactone of a water-soluble organic acid, and other compounds of similarfunction well known to those trained in the art. The nasal formulationsmay also be in the form of an aerosol comprising the peptide togetherwith an extender which may be an amino acid, for example, methionine.

Transdermal application formulations can comprise the peptides and theirlipid conjugates, optionally incorporated into a suitable topicalcarrier, or a dermal patch. The compounds can be combined with apenetration-enhancing agent, for example, dimethylsulfoxide (DMSO),dimethylformamide, dimethylacetamide, or Azone®, azacycloheptane-2-one.Both topical carriers and the use of penetration enhancers are disclosedby Blaug, S., Chap. 87 Remington's Pharmaceutical Sciences, 15th Ed.Mack Publishing Co., Easton, Pa. 18042 (1975).

In the therapy of HIV-infected patients, the lipid conjugated proteaseinhibiting peptide will be administered parenterally, orally, ornasally. Parenteral doses are usually from 0.01 to 1 gm every 3-8 hours.Nasal or oral doses may be from 2 to 10 times the parenteral dose,depending on bioavailability of the peptide, and its retention in theplasma and tissues. The most efficacious dose can be readily determinedby standard pharmacokinetic and toxicological studies in animals andhumans.

EXPERIMENTAL PROCEDURES

The amino acid abbreviations used are those commonly employed in thepeptide art and described in the literature, eg. IUPAC-IUB Commission onBiochemical Nomenclature, J. Biol. Chem. 247, 979-982 (1972). Additionalabbreviations used are: Ac=Acetyl; Boc=t-Butyloxycarbonyl; Suc-succinicacid; --OMe=Methyl ester, --NH₂ =Amide; --NHOH=Hydroxylamide; --NHNH₂=Hydrazide; DPPA=1,2-dipalmitoylphosphatidic acid;DPPE=1,2-dipalmitoyl-phosphatidyl ethanolamine; Achx=1-Amino,1-cyclohexane carboxylic acid; Acpr=1-Amino, 1-cyclopropane carboxylicacid; Acpnt=1-Amino, 1-cyclopentane carboxylic acid; Pip=Pipecolic acid(4-piperidine carboxylic acid); Ava=5-Amino valerie acid,a-(OH)-Leu=L-Leucic acid (2-OH-L-isocaproic acid); Nal=Naphthylalanine;Phe(4-Cl)=p-Chloro phenylalanine; Nva=norvaline; Paa=Phosphonoaceticacid; Chl=Cholic acid. 3,5-di-Aba=3,5-diaminobenzoic acid. Amino acidsdiscussed herein are of the L-form unless otherwise mentioned.

All the temperatures described are in degree Celsius and areuncorrected. Evaporations were carried out under vacuum below 35° C. TLCwas carried out using E.Merck precoated plates and the spots werevisualized by exposure to UV light, iodine, ninhydrin spray, phosphorousspray, or sulfuric acid spray followed by charring as appropriate.Analytical HPLC was carried out using a Beckman system and Vydac reversephase columns (C-4 or C-18 as appropriate). Preparative HPLC was carriedout using a Waters Deltaprep system using either reverse phase or silicacolumns. Purity and authenticity of the compounds were established byTLC, analytical HPLC, amino acid analysis, elemental analysis, UVspectra, NMR, FAB-MS as required and appropriate.

The chemical reactions described below are generally disclosed in termsof their general application to the preparation of peptides of theinvention. Occasionally, the reaction may not be applicable as describedto each peptide included within the disclosed scope. The peptides forwhich this occurs will be readily recognized by those skilled in theart. In all such cases, either the reactions can be successfullyperformed by conventional modifications known to those skilled in theart, e.g. by appropriate protection of interfering groups, by changingto alternative conventional reagents, or by routine modification ofreaction conditions. Alternatively, other reactions disclosed herein orotherwise conventional will be applicable to the preparation of thecorresponding peptides of the invention. In all preparative methods, allstarting materials are known or readily preparable from known startingmaterials.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the invention to itsfullest extent. The following preferred embodiments are, therefore, tobe construed as merely illustrative and not limitative of the remainderof the disclosure in any way whatsoever. In the following examples, alltemperatures are set forth uncorrected in degrees Celsius; unlessotherwise indicated, all parts and percentages are by weight.

The present invention is described below in detail using the followingexamples, but the methods described below are applicable for thepreparation of all peptides covered by the scope of the invention andare not limited to the Examples given below.

EXAMPLE 1

General procedure for coupling phosphatidic acid to the amino group of apeptide

1,2-Dipalmitoyl-sn-glycero-3-phosphatidic acid.2Na (DPPA.2Na; AvantiPolar Lipids, Birmingham Ala.; MW: 697.84; 698 mg, 1 mmol) waspartitioned between chloroform:methanol (2:1 (v/v); 200 ml) and cold 1NHCl (50 ml). The aqueous layer was re-extracted with chloroform methanol(2:1 (v/v); 100 ml). The combined organic phase was evaporated and driedunder vacuum over P₂ O₅. The resulting free phosphatidic acid wasdissolved in a mixture of DMF (2 ml) and pyridine (2 ml) and to thesolution were added the appropriate peptide having a free amino group (1mmol) followed by N,N'-dicyclohexylcarbodiimide (DCC; Aldrich ChemicalCo. Milwaukee, Wis., MW: 206, 620 mg, 3 mmol). The reaction mixture wasstirred for 24 hours at room temperature. The solvents were evaporatedand the product was purified by flash chromatography over silica gelcolumn (2.5×50 cm) using a linear gradient of 0 to 50% methanol inchloroform. Fractions containing the desired product as indicated by TLCand HPLC were pooled and evaporated. The product was further purified,if necessary, by preparative HPLC or by crystallization.

EXAMPLE 1A

A: DPPA-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂ and

EXAMPLE 1B

DPPA-Ser-Gln-Asn-Tyr-Acpnt-Ile-Val-NH₂ peptides were prepared by theabove procedure.

EXAMPLE 2

General procedure for coupling phosphatidic acid to the hydroxy group ofa peptide

1,2-Dipalmitoyl-sn-glycero-3-phosphatidic acid (1 mmol) prepared asabove was dissolved in a mixture of DMF (2 ml) and pyridine (2 ml) andto the solution were added the appropriate peptide having a freehydroxyl group (1 mmol) followed by DCC (620 mg, 3 mmol). The reactionwas carried out and the product was isolated as described in Example 1.

The condensation of the phosphatidic acid and the hydroxyl group of apeptide was also conveniently carried out by using2,4,6-triisopropylbenzenesulfonyl chloride (TPS-Cl; Aldrich ChemicalCo., Milwaukee, Wis.; MW: 302.86; 758 mg, 2.5 mmol) as a coupling agentin place of DCC.

EXAMPLE 3

General procedure for coupling a peptide containing a free carboxylgroup to the amino group of a phosphatidylethanolamine

A mixture of the appropriate peptide (1 mmol), phosphatidyl-ethanolamine(1 mmol) were dissolved in pyridine (5 ml) and DCC (3 mmol) followed by1-hydroxybenzotriazole (HOBt; Aldrich Chemical Co., HOBt, MW: 153; 450mg, 3 mmol) were added. The reaction mixture was stirred for 24 hours atroom temperature and the product was purified by silica gelchromatography as described in Example 1.

EXAMPLE 3A

i-Boc-Tyr-Pro-Ile-Gly-DPPE was prepared by the above procedure.

EXAMPLE 4

General procedure for coupling peptide containing a free carboxyl groupto the hydroxy group of a diacyl or dialkyl glycerol

A mixture of the appropriate peptide (1 mmol), diacyl or dialkylglycerol (1 mmol) were dissolved in pyridine (5 ml) and DCC (3 mmol)followed by 4-dimethylaminopyridine (Aldrich Chemical Co., DMAP, MW:122.17; 122 mg, 1 mmol) were added. The reaction mixture was stirred for24 hours at room temperature and the product was purified by silica gelchromatography as described in Example 1.

EXAMPLE 5

Preparation of N-tert.Butyloxycarbonyl-Serine-Hydroxysuccinimide Ester(Boc-Ser-OSu) MW: 302.26

Boc-Ser-OH (20.52 g, 100 mmol) and N-hydroxysuccinimide (AldrichChemical Co., Milwaukee, Wis.; MW: 115.09; 23.3 g, 200 mmol) weredissolved in dry THF (400 ml). The solution was cooled to -10° C., DCC(MW: 206.33; 30.10 g; 150 mmol) was added. The mixture was stirred for 2hours at -10° C. and overnight at room temperature. The solvents wereevaporated under vacuum, and the residue was stirred in ethyl acetate (1l) and filtered to remove insoluble material. The resulting filtrate waswashed successively with water (3×250 ml), cold 1N HCl (3×250 ml), water(3×250 ml), cold 10% NaHCO₃ (3×250 ml) and water (3×250 ml) and driedover anhydrous sodium sulfate. After evaporation the residue wascrystallized from isopropyl alcohol.

Boc-Tyr-OSu, Boc-Hyp-OSu and Boc-Thr-OSu were prepared similarly.

EXAMPLE 6

Preparation of 1,2-Dipalmitoyl-sn-glycero-3-phospho-O-(N-Boc)-Ser-OSu;Boc-Ser(ODPP)-OSu: MW: 850.17

1,2-Dipalmitoyl-sn-glycero-3-phosphatidic acid (1 mmol) was dissolved inanhydrous pyridine (5 ml) and 2,4,6-triisopropylbenzenesulfonyl chloride(TPS-Cl, 758 mg, 2.5 mmol) followed by Boc-Ser-OSu (MW: 302.26; 302 mg,1 mmol) was added. The reaction mixture was stirred under dry nitrogenatmosphere for 16 hours at room temperature. The reaction was thenquenched by adding water (1 ml) and the solvents were evaporated undervacuum. The residue was dissolved in chloroform (5 ml) and loaded onto asilica gel 60 column (2.5 cm×45 cm) equilibrated with chloroform. Thecolumn was eluted with a gradient of chloroform (500 ml) to 15% MeOH inchloroform (500 ml). Fractions containing the desired product (asindicated by TLC) were pooled and evaporated to furnishBoc-Ser(ODPP)-OSu.

Boc-Tyr(ODPP)-OSu, Boc-Hyp(ODPP)-OSu and Boc-Thr(ODPP)-OSu were preparedsimilarly.

EXAMPLE 7

General procedure for coupling phosphatidyl-serine, to a peptidecontaining a free amine function.

Boc-Ser(ODPP)-OSu (0.1 mmol) and the peptide in question containing afree amine function (0.1 mmol) were dissolved in DMF (2 ml) and pyridine(2 ml) and stirred for 24 hours at room temperature. The solvents wereevaporated and the product was purified by flash chromatography oversilica gel column (2.5×50 cm) using a linear gradient of 0 to 50%methanol in chloroform. Fractions containing the desired product asindicated by TLC and HPLC were pooled and evaporated. The product wasfurther purified, if necessary, by preparative HPLC or bycrystallization and deprotected by treatment with trifluoroacetic acid(2 ml) for 12 hours at room temperature. The acid was removed byevaporation under vacuum and the product was isolated by lyophilizationfrom water.

Peptides linked to phosphatidyl-tyrosine, phosphatidyl-hydroxyprolineand phosphatidyl-threonine were prepared similarly.

EXAMPLE 7A ##STR2## according to the above procedure. EXAMPLE 8

Preparation of 1,2-Dipalmitoyl-sn-glycero-3-phospho-O-(N-Fmoc)-Ser-OBzl(Fmoc-Ser(ODPP)-OBzl)

Starting from Fmoc-Ser-OBzl the title compound was prepared followingthe procedure described for Boc-Ser(ODPP)-OSu.

Fmoc-Tyr (ODPP)-OBzl, Fmoc-Hyp(ODPP)-OBzl and Fmoc-Thr(ODPP)OBzl wereprepared similarly.

EXAMPLE 9

General procedure for coupling a peptide containing a free carboxylgroup to phosphatidyl-serine

Fmoc-Ser(ODPP)-OBzl (0.1 mmol) was dissolved in DMF (2 ml) andpiperidine (0.2 ml) and the mixture was stirred under dry nitrogenatmosphere for 4 hours and the solvents were evaporated under vacuum.The residue was dissolved in a dry DMF (5 ml) and the peptide inquestion containing a free carboxyl group (0.1 mmol) was added to itfollowed by DCC (0.1 mmol) and HOBt (0.1 mmol). The reaction mixture wasstirred for 24 hours at room temperature. The solvents were evaporatedand the product was purified by flash chromatography over silica gelcolumn (2.5×50 cm) using a linear gradient of 0 to 50% methanol inchloroform. Fractions containing the desired product as indicated by TLCand HPLC were pooled and evaporated. The product was further purified,if necessary, by preparative HPLC or by crystallization. The product wasthen dissolved in DMF (5 ml) and hydrogenated in the presence ofpalladium-carbon (5%) catalyst (200 mg). The catalyst was removed byfiltration and the product was purified by silica gel chromatography asdescribed above.

Peptides containing phosphatidyl-tyrosine, phosphatidyl-hydroxyprolineand phosphatidyl-threonine were prepared similarly.

EXAMPLE 10

Preparation ofBoc-Serine(O-diphosphate-dipalmitoylglycerol)-N-hydroxysuccinimide ester(Boc-Ser(O-DP-DPG)-ONSu)

A. Boc-Serine(O-phosphate)-N-hydroxysuccinimide ester

Boc-Ser-OSu (MW: 302.26; 302 mg, 1 mmol) was dissolved in triethylphosphate (5 ml) and cooled to -5° C. POCl₃ (MW 153.33; 460 mg, 3 mmol)was added to the above solution and the reaction mixture was stirredunder dry nitrogen atmosphere for 16 hours at -10° C. Ether (20 ml) wasadded and the resulting precipitate was separated from the supernatantby decantation and the residue was washed with ether (20 ml each). Theresidue was resuspended in ice cold water (10 ml) and the pH wasimmediately adjusted to 7.5 by adding 1N NaOH. The solution was stirredfor 1 hour at 0° C. while maintaining the pH at 7.5 and lyophilized. Theresulting product was dissolved in water (5 ml) and loaded onto a DEAESephadex A-25 column (1 cm×10 cm) equilibrated with 0.01M ammoniumbicarbonate (pH 7). The column was eluted with a gradient of 0.01Mammonium bicarbonate to 0.3M ammonium bicarbonate (pH 7). Fractionscontaining the desired product (as indicated by TLC) were pooled andlyophilized to furnish Boc-Ser(O-phosphate)-Osu.

B. 1,2-Dipalmitoyl-sn-glycero-3-phosphoromorpholidate(DPPA-Morpholidate)

1,2-Dipalmitoyl-sn-glycero-3-phosphatidic acid (DPPA, 650 mg, 1 mmol)and morpholine (350 mg, 4 mmol) were dissolved in chloroform (10 ml),t-butanol (10 ml) and water (1 ml). The solution was stirred undergentle reflux for 2 hours while adding a solution of DCC (825 mg, 4mmol) in t-butanol (10 ml). The mixture was stirred for an additional 4hours and evaporated to dryness and suspended in water (100 ml) and wasextracted three times with ether (100 ml), evaporated to dryness andlyophilized.

C. Coupling of DPPA-Morpholidate and Boc-Ser(O-phosphate)-OSu

DPPA-Morpholidate (0.5 mmol) and Boc-Ser(O-phosphate)-OSu (0.3 mmol)were dissolved in anhydrous pyridine (25 ml). The solution wasevaporated to dryness in vacuo five times from anhydrous pyridine,followed by the addition of anhydrous pyridine (5 ml). The reactionmixture was stirred at 40° C. and evaporated to dryness. The product wasdissolved in chloroform:methanol:water (2:3:1) and loaded onto a DEAESephadex A-25 column (1 cm×10 cm) equilibrated withchloroform:methanol:water (2:3:1). The column was eluted with a lineargradient of 0 to 0.3M ammonium bicarbonate in the same solvent.Fractions containing the desired product (as indicated by TLC) werepooled, concentrated to a 60 ml volume and extracted 5 times withchloroform (50 ml each). The cloroform solution was evaporated tofurnish Boc-Serine(O-diphosphatedipalmitoylglycerol)-N-hydroxysuccinimide ester.

Boc-Tyr(O-DP-DPG)-ONSu, Boc-Hyp(O-DP-DPG)-ONSu andBoc-Thr(O-DP-DPG)-ONSu were prepared similarly.

EXAMPLE 11

General procedure for coupling Serine-O-diphosphate dipalmitoylglycerolto a peptide containing a free amine function.

Boc-Ser(O-DP-DPG)-ONSu (0.1 mmol) and the peptide in question containinga free amine function (0.1 mmol) were dissolved in DMF (2 ml) andpyridine (2 ml) and stirred for 24 hours at room temperature. Thesolvents were evaporated and the product was purified by flashchromatography over silica gel column (2.5×50 cm) using a lineargradient of 0 to 50% methanol in chloroform. Fractions containing thedesired product as indicated by TLC and HPLC were pooled and evaporated.The product was further purified, if necessary, by preparative HPLC orby crystallization and deprotected by treatment with trifluoroaceticacid (2 ml) for 12 hours at room temperature. The acid was removed byevaporation under vacuum and the product was isolated by lyophilizationfrom water.

Tyrosine-O-diphosphate dipalmitoylglycerol, hydroxyproline-O-diphosphatedipalmitoylglycerol and threonine-O-diphosphate dipalmitoylglycerolcontaining peptides were prepared similarly.

EXAMPLE 12

Fmoc-Serine(O-diphosphate dipalmitoylglycerol)-Benzyl Ester(Fmoc-Ser(O-DP-DPG)-OBzl)

Starting from Fmoc-Ser-OBzl and DPPA-Morpholidate the title compound wasprepared following the procedure described in Example 11.

Fmoc-Tyr(O-DP-DPG)-OBzl, Fmoc-Hyp(O-DP-DPG)-OBzl andFmoc-Thr(O-DP-DPG)-OBzl were prepared similarly.

EXAMPLE 13

General procedure for coupling a peptide containing a free carboxylgroup to Serine-O-diphosphate dipalmitoylglycerol

Starting from Fmoc-Ser(O-DP-DPG)-OBzl, the title compound was preparedby deprotection of the Fmoc- group, followed by the coupling reaction,the catalytic hydrogenation reaction and the purification proceduredescribed above.

Corresponding tyrosine-O-diphosphate dipalmitoylglycerol,hydroxyproline-O-diphosphate dipalmitoylglycerol andthreonine-O-diphosphate dipalmitoylglycerol containing peptides wereprepared similarly.

EXAMPLE 14

Preparation of 1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Ser-OSu

Boc-Ser(O-phosphate)-OSu (1 mmol) was dissolved in anhydrous pyridine (5ml) and 2,4,6-triisopropylbenzenesulfonyl chloride (TPS-Cl; AldrichChemical Co., Milwaukee, Wis.; MW: 302.86; 758 mg, 2.5 mmol) was addedfollowed by 1-O-Hexadecyl-sn-glycerol. The reaction mixture was stirredunder dry nitrogen atmosphere for 16 hours at room temperature. Thereaction was then quenched by adding water (1 ml) and the solvents wereevaporated under vacuum. The residue was dissolved in chloroform (5 ml)and loaded onto a silica gel 60 column (2.5 cm×45 cm) equilibrated withchloroform. The column was eluted with a gradient of chloroform (500 ml)to 15% MeOH in chloroform (500 ml). Fractions containing the desiredproduct (as indicated by TLC) were pooled and evaporated to furnish thetitle compound.

Preparation of 1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Tyr-Osu,1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Hyp-OSu and1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Thr-OSu were carried out ina similar manner.

EXAMPLE 15 General procedure for coupling1-O-hexadecyl-sn-glycero-3-phospho-serine to a peptide containing freeamine function

This coupling reaction, subsequent deprotection and purification of theproduct was carried out following the procedure described in Example 9except for using 1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Ser-OSu inplace of Boc-Ser(ODPP)-OSu.

Peptides containing 1-O-hexadecyl-sn-glycero-3-phospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-phospho-O-hydroxyproline and1-O-hexadecyl-sn-glycero-3-phospho-O-threonine were prepared in asimilar manner.

EXAMPLE 16

Preparation of 1-O-hexadecyl-sn-glycero-3-phospho-O-(Fmoc)-Ser-OBzl

Starting from Fmoc-Ser-OBzl the title compound was prepared followingthe procedure described for the preparation of1-O-hexadecyl-sn-glycero-3-phospho-O-(N-Boc)-Ser-OSu.

Preparation of 1-O-hexadecyl-sn-glycero-3-phospho-O-(Fmoc)-Tyr-OBzl,1-O-hexadecyl-sn-glycero-3-phospho-O-(Fmoc)-Hyp-OBzl and1-O-hexadecyl-sn-glycero-3-phospho-O-(Fmoc)-Thr-OBzl were carried outsimilarly.

EXAMPLE 17

General procedure for coupling a peptide containing free carboxyl groupto 1-O-hexadecyl-sn-glycero-3-phospho-serine

Starting from 1-O-hexadecyl-sn-glycero-3-phospho-O-(Fmoc)-Ser-OBzl,deprotection of the Fmoc- group, the coupling reaction, catalytichydrogenation followed by purification as described in Example 10.

Peptides containing 1-O-hexadecyl-sn-glycero-3-phosphotyrosine,1-O-hexadecyl-sn-glycero-3-phospho-hydroxyproline and1-O-hexadecyl-sn-glycero-3-phospho-threonine were prepared in a similarmanner.

EXAMPLE 18

Preparation of 1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Ser-OSu

Boc-Ser(O-phosphate)-OSu (1 mmol) was converted to the correspondingmorpholidate by reacting with DCC and morpholine. The product was thenreacted with 1-O-hexadecylglycerol-3-phosphate as described for thepreparation of Boc-Ser(O-DP-DPG)-OSu.

Preparation of 1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Tyr-OSu,1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Hyp-OSu and1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Thr-OSu were carried outin a similar manner.

EXAMPLE 19

General procedure for coupling1-O-hexadecyl-sn-glycero-3-diphospho-serine to a peptide containing freeamine function

This coupling reaction, subsequent deprotection and purification of theproduct was carried out following the procedure described in Example 9except for using 1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Ser-OSuin place of Boc-Ser(O-DP-DPG)-OSu.

Peptides containing 1-O-hexadecyl-sn-glycero-3-diphospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-diphospho-O-hydroxyproline and1-O-hexadecyl-sn-glycero-3-diphospho-O-threonine were prepared in asimilar manner.

EXAMPLE 20

Preparation of 1-O-hexadecyl-sn-glycero-3-diphospho-O-(Fmoc)-Ser-OBzl

Starting from Fmoc-Ser-OBzl the title compound was prepared followingthe procedure described for the preparation of1-O-hexadecyl-sn-glycero-3-diphospho-O-(N-Boc)-Ser-OSu.

Preparation of1-O-hexadecyl-sn-glycero-3-diphospho-O-(Fmoc)-Tyr-OBzl,1-O-hexadecyl-sn-glycero-3-diphospho-O-(Fmoc)-Hyp-OBzland 1-O-hexadecyl-sn-glycero-3-diphospho-O-(Fmoc)-Thr-OBzl were carriedout in a similar manner.

EXAMPLE 21 General procedure for coupling a peptide containing freecarboxyl group to 1-O-hexadecyl-sn-glycero-3-diphospho-serine

Starting from 1-O-hexadecyl-sn-glycero-3-diphospho-O-(Fmoc)-Ser-OBzl,the title compound was obtained by deprotection of the Fmoc- group,followed by the coupling reaction, the catalytic hydrogenation and thepurification procedure described in Example 9.

Peptides containing 1-O-hexadecyl-sn-glycero-3-diphospho-O-tyrosine,1-O-hexadecyl-sn-glycero-3-diphospho-O-hydroxyproline and1-O-hexadecyl-sn-glycero-3-diphospho-O-threonine were prepared in asimilar manner.

EXAMPLE 22

Preparation of1,2-Dipalmitoyl-sn-glycero-3-phospho-O-(N-Succinyl)-ethanolamineDPPE-Succinic Acid MW: 791.03

To a solution of 1,2-Dipalmitoyl-sn-glycero-3-phosphatidyl ethanolamine(DPPE; Avanti Polar Lipids, Birmingham, Ala.; MW: 690.96; 346 mg, 0.5mmol) in chloroform (10 ml), succinic anhydride (Sigma Chemical Co., St.Louis, Mo.; MW: 100.7, 100 mg, 1 mmol) dissolved in chloroform (10 ml)and triethylamine (0.1 ml) were added. The reaction mixture was stirredunder dry nitrogen atmosphere for 16 hours at room temperature. Thereaction was monitored by running TLC at various intervals. After thereaction is completed, the solvents were evaporated under vacuum and theresidue was dissolved in chloroform (5 ml) and loaded onto a silica gel60 column (2.5 cm×45 cm) equilibrated with chloroform. The column waseluted with a gradient of chloroform (500 ml) to 15% MeOH in chloroform(500 ml). Fractions containing the desired product (as indicated by TLC)were pooled and evaporated to furnish DPPE-Succinic Acid ACID.

Preparation of1,2-Dimyristoyl-sn-glycero-3-phospho-O-(N-Succinyl)-ethanolamine,DMPE-SUCCINIC ACID, was carried out similarly.

EXAMPLE 23

Preparation of ##STR3##

A mixture of Boc-Phe-OH (5.3 g, 20 mmol), triethylamine (TEA; MW:101.19; 3.1 g, 31 mmol), anhydrous KF (1.2 g, 20 mmol) and1,3-dibromobutane-2,3-diol (Br-CH₂ --CHOH--CHOH-CH₂ --Br, 2.5 g, 10mmol) was dissolved in DMF (25 ml) and the solution was stirred for 24hr at 45° C. The solvent was evaporated under vacuum and residue wasdissolved in EtAc (330 ml), extracted with 10% sodium bicarbonate (3×50ml), water (3×50 ml), 10% citric acid (3×50 ml) and water (3×50 ml). Theorganic phase was dried over anhydrous sodium sulphate and evaporatedunder vacuum and the residue was crystallized from acetonitrile tofurnish the product as white amorphous powder.

EXAMPLE 24

Preparation of ##STR4##

The compound from example 23 (616 mg, 1 mmol) was dissolved in anhydrousTFA (5 ml) and stirred for 15 minutes. The solvent was evaporated andthe residue triturated with ether and filtered. The product wasdissolved in DMF (20 ml) and cooled to -10° C. Boc-Val-OH (MW: 217.14,696 mg, 3 mmol), HOBt (456 mg, 3 mmol) and DCC (620 mg, 3 mmol) wereadded to the above solution. The reaction mixture was stirred for 2hours at -10° C. and for 16 hours at room temperature. The solvents werethen evaporated under vacuum, and the residue was dissolved in ethylacetate (250 ml) and filtered to remove insoluble material. Theresulting filtrate was washed successively with water (3×50 ml), cold 1NHCl (3×50 ml), water (3×50 ml), cold 10% NaHCO₃ (3×50 ml) and water(3×50 ml) and dried over anhydrous sodium sulfate. Evaporation of thesolvent provided the crude product which was purified by crystallizationfrom acetonitrile.

EXAMPLE 25

Preparation of ##STR5##

The title compound was prepared following the procedure described forExample 24 except using Boc-Ser-OH instead of Boc-Val-OH.

EXAMPLE 26

Preparation of ##STR6##

The title compound was prepared following the procedure described forExample 24 except using Boc-Asn-OH instead of Boc-Val-OH.

EXAMPLE 27

General method for the synthesis of peptides by the solid phase method.

A. Resin Peptide Synthesis

Boc-amino acyl-benzyl ester resin or Boc-aminoacyl-(4-methyl)benzhydrylamine resin (Boc-AA-Resin, 2 g, 1 mmol) placedin a reaction vessel of Beckman 990 B Peptide Synthesizer (BeckmanInstruments, Palo Alto, Calif.) and subjected to the followingoperations. Each step is carried out one time unless specifiedotherwise, and reagents and solvents after each step are separated fromthe peptide resin by filtration under nitrogen.

    ______________________________________                                                                      Mix Time                                        Step  Reagent/solvent/No of times                                                                           (minutes)                                       ______________________________________                                        1     DCM (30 ml, 3 times)    1.5                                             2     TFA-DCM (1:1) (30 ml)   1.5                                             3     TFA-DCM (1:1) (30 ml)   30.0                                            4     DCM (30 ml, 3 times)    1.5                                             5     Methanol (30 ml, 3 times)                                                                             1.5                                             6     DCM (30 ml, 3 times)    1.5                                             7     TFA-DCM (1:1) (30 ml)   1.5                                             8     TFA-DCM (1:1) (30 ml)   5.0                                             9     DCM (30 ml, 3 times)    1.5                                             10    DMF (30 ml, 3 times)    1.5                                             11    Boc--Thr(Bzl)--OH/HOBt/DCC                                                                            240.0*                                                (4 mmol each) in DMF (20 ml)                                            12    DCM (30 ml, 3 times)    1.5                                             13    Methanol (30 ml, 3 times)                                                                             1.5                                             14    DCM (30 ml, 3 times)    1.5                                             ______________________________________                                         *Coupling reaction was carried out for an average of 4 hours, as in this      case, or until a ninhydrin test (Kaiser E. T. et al, Anal. Biochem. 34,       595-8, 1969) showed a negative result indicating the absence of free amin     groups. The same sequence of reactions was repeated using appropriate         amino acid derivatives until the required peptide chain was assembled on      the resin. After completion of the synthesis, the resin was removed from      the vessel and dried under vacuum.                                       

B. Cleavage of the Resin-Peptide using Hydrogen Fluoride (HF):

Peptides containing free carboxyl groups or carboxamide function at theC-terminal are prepared by treating the corresponding benzyl esterlinked or 4-methylbenzhydrylamine linked peptide resins according to thefollowing procedure. The dried peptide resin (1 g), anisole (1 ml) andp-cresol (0.1 g) were placed in a Kel F reaction vessel. The vessel wasplaced in a bath of liquid nitrogen and anhydrous HF (15 ml) wascondensed into the vessel. The reaction mixture was stirred at -10° C.for 1 hour and HF was removed by evaporation under vacuum. The residuewas triturated with dry ether (50 ml), filtered and washed withadditional quantity of ether (3×50 ml). Peptide product in the mixturewas isolated by extracting with glacial acetic acid (3×50 ml) followedby lyophilization of the solvent.

C. Preparation of peptide methyl esters by transesterification:

Boc-peptidyl-benzyl ester resin (1 g) was stirred with methanol (10 ml)and triethylamine (1 ml) for 18 hours at room temperature. The resin wasfiltered and washed three times with methanol (20 ml each time) and thecombined filtrate was evaporated to provide peptide methyl esters. Someof the peptides prepared by this method contain protecting groups forother functional groups which are very conveniently deprotected bytreatment with liquid HF as described above. The resulting products werepurified as described below.

D. Preparation of peptide hydrazides:

Boc-peptidyl-benzyl ester resin (1 g) was stirred with methanol (10 ml)and anhydrous hydrazine (1 ml) for 18 hours at room temperature. Theresin was filtered and washed three times with methanol (20 ml eachtime) and the combined filtrate was evaporated to provide peptidehydrazides esters. Some of the peptides prepared by this method containprotecting groups for other functional groups which are veryconveniently deprotected by treatment with liquid HF as described above.The resulting products were purified as described below.

E. Peptide Purification:

Peptide powder obtained above (200 mg) was dissolved in 1N acetic acid(3 ml), loaded to a Sephadex G-25 (superfine) column (1.5 cm×100 cm) andeluted with 1N acetic acid. The eluent fractions containing the peptidewere pooled and freeze dried. The resulting peptide (50 mg) was furtherpurified by preparative reverse phase high performance chromatography(RP-HPLC) using a Waters C-4 column and a buffer gradient of 0.1% TFA inwater to 70% acetonitrile in 0.1% TFA in water. The fractions containingpure peptide (determined by analytical HPLC) were combined and theproduct isolated by lyophilization. Purity of the peptide was betterthan 95% by HPLC; and amino acid analysis followed by acid hydrolysis(6N HCl, 110° C., 24 hr) gave expected amino acid ratios.

Peptides of the foregoing examples listed below were prepared by thesolid phase method.

EXAMPLE 27-01 Ser-Gln-Asn-Phe-Pro-Ile-Val-NH₂

EXAMPLE 27-02 Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂

EXAMPLE 27-03 Ser-Gln-Asn-Tyr-Achx-Ile-Val-NH₂

EXAMPLE 27-04 Ser-Gln-Asn-Tyr-Acpr-Ile-Val-NH₂

EXAMPLE 27-05 Ser-Gln-Asn-Tyr-Acpnt-Ile-Val-NH₂

EXAMPLE 27-06 Thr-Ile-Leu-(beta-Ala)-Leu-Gln-Arg-NH₂

EXAMPLE 27-07 Ser-Gln-Asn-Tyr-Pro-Ile-Val-Thr-Leu-Ava-Thr-Gln-Arg-NH₂

EXAMPLE 27-15 Ac-Ala-Ala-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-16 Ac-Ala-Ala-Phe-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-17 Ac-Ala-Ala-(DL-Phe(4-Cl))-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-18 Ala-Ala-Phe-(beta-Ala)-Val-Val-Gly-OH

EXAMPLE 27-19 Ala-Ala-Phe-(beta-Ala)-Nva-Val-Gly-OH

EXAMPLE 27-20 Ala-Ala-Phe-(beta-Ala)-(a-(OH)-iso-Valeryl)-Val-GLy-OH

EXAMPLE 27-21 Ala-Ala-Phe-(beta-Ala)-Val-Val-Gly-OMe

EXAMPLE 27-22 Ala-Ala-Phe-(beta-Ala)-Nva-Val-Gly-OMe

EXAMPLE 27-23 Ala-Ala-Phe-(beta-Ala)-(a-(OH)-iso-Valeryl)-Val-Gly-OMe

EXAMPLE 27-24 Boc-Ala-Ala-Phe-(beta-Ala)-Nva-Gly-OMe

EXAMPLE 27-25 (a,e-dimyristoyl-K)-dP-Ava-Ala-Ala-Phe-Ava-Val-Gly-OMe

EXAMPLE 27-26 Ac-Ala-Ala-(D-a-Nal)-Pip-OMe

EXAMPLE 27-27 Ac-Ala-Ala-(D-a-Nal)-Pip-NHNH₂

EXAMPLE 27-28 Ac-Ala-Ala-(L-a-Nal)-Pip-OMe

EXAMPLE 27-29 Ac-Ala-Ala-(L-a-Nal)-Pip-NHNH₂

EXAMPLE 27-30 Ac-Ala-Ala-(D-b-Nal)-Pip-OMe

EXAMPLE 27-31 Ac-Ala-Ala-(D-b-Nal)-Pip-NHNH₂

EXAMPLE 27-32 Chl-Ala-Ala-(L-a-Nal)-Pip-OMe

EXAMPLE 27-33 Chl-Ala-Ala-(L-a-Nal)-Pip-NHNH₂

EXAMPLE 27-34 Paa-Ala-Ala-(D-a-Nal)-Pip-OMe

EXAMPLE 27-36 Ac-Ala-Ala-(DL-Phe(4-Cl))-Pip-OH

EXAMPLE 27-38 Val-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂

EXAMPLE 27-39 Val-Ser-Gln-Asn-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-40 i-Boc-(D-Phe)-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-41 (i-Boc-(D-Phe)-Leu-)₂ -(3,5-di-Aba)-Pro-Leu-(D-Phe)-NH₂

EXAMPLE 27-42 (Myristoyl-(D-Phe)-Leu-)₂-(3,5-di-Aba)-Pro-Leu-(D-Phe)-NH₂

EXAMPLE 27-43 i-Boc-(3,5-di-Aba)-(D-a-Nal)-Pip-(a-(OH)-Leu)-Val-NH₂

EXAMPLE 27-46 Pro-Ile-Val-NH₂

EXAMPLE 27-47 Ser-Gln-Asn-Tyr

EXAMPLE 27-48 Asn-Phe(CO--CH₂ N)Pip-Ile-NH₂

EXAMPLE 27-49 Quinaldoyl-Asn-Phe(CO--CH₂ N)Pip-Ile-NH₂

EXAMPLE 27-50 Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂

EXAMPLE 27-51 Quinaldoyl-Asn-Phe(CHOH--CH₂ N)Diq-Ile-Val-Gln-NH₂

EXAMPLE 27-52 Val-Ser-Gln-Asn-Tyr-Diq-Ile-Val-Gln-NH₂

EXAMPLE 27-53 Boc-Asn-Phe(CO--CH₂ N)Diq-NHtBu

EXAMPLE 27-54 Succinyl-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂

EXAMPLE 27-55 Succinyl-Ser-Gln-Asn-Tyr

EXAMPLE 27-56 Quinaldoyl-Asn-Phe(CHOH--CH₂ N)Pip-Ile-NH₂

EXAMPLE 27-60 Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂

EXAMPLE 27-61 Phe(CHOH--CH₂ N)Pro-Ile-Val-NH₂

EXAMPLE 27-62 His-Lys-Arg-Ala-Val-Leu-Phe(4-NO₂)-Glu-Ala-Nle-Ser-NH₂

EXAMPLE 27-69 [D-Phe]-[D-a-Nal]-Pip-[L-a-(OH)-Leu]-Val-NH₂

EXAMPLE 27-70 Boc-Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂

EXAMPLE 27-71 Boc-Phe(CHOH--CH₂ N)Pro-Ile-Val-NH₂

EXAMPLE 27-74 Succinyl-Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂

EXAMPLE 27-77 [D-Phe]-[D-b-Nal]-Pip-[L-a-(OH)-Leu]-Val-NH₂

EXAMPLE 27-79 i-Boc-Tyr-Pro-Ile-Gly-OH

EXAMPLE 28

General procedure for the coupling of DMPE-Succinic Acid orDMPE-Succinic acid to peptides containing free amine group

DPPE-SUCCINIC ACID or DMPE-Succinic Acid (1 mmol) and the requiredpeptide having a free amine function (1 mmol) were dissolved in DMF (5ml) and DCC (3 mmol) followed by 1-hydroxybenzotriazole (HOBt; AldrichChemical Co., HOBt, MW: 153; 450 mg, 3 mmol) were added. The reactionmixture was stirred for 24 hours at room temperature and the product waspurified by silica gel chromatography as described in Example 1.

This procedure was used to prepare the following peptides:

EXAMPLE 28A: DPPE-Suc-Ala-Ala-(D-b-Nal)-Pip-OMe

EXAMPLE 28B: DPPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂

EXAMPLE 28C: DMPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂

EXAMPLE 28D: DMPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Diq-Ile-Val-Gln-NH₂

EXAMPLE 28E: DPPE-Succinyl-[D-Phe]-[D-a-Nal]-Pip-[L-a-(OH)-Leu]-Val-NH₂

EXAMPLE 28F: DPPE-Succinyl-Phe(CHOH--CH₂ N)Phe-Ile-Phe-NH₂

EXAMPLE 28G: DPPE-Succinyl-Phe-O--CH₂ --CHOH NH₂ -Phe-O--CH₂ --CHOH

EXAMPLE 29

Assay For The Reduction of HIV Syncytia Formed In HT4-6C Cells

The syncytia reduction assay as described by Larder, B. et al., science243:1731-1734(1989) was used to measure the antiviral effect of proteaseinhibitor. A Hela cell line, HT4-6C, expressing the human CD4 receptoron its surface was trypsinized with 0.25% trypsin for 5 min. Cells werecentrifuged to remove the residual trypsin, and the cell pellet wasresuspended in DMEM with 10% FCS. HeLa cells were plated in 96 wellplate (1×10⁵ cells/well) overnight. Cell cultures were infected with HIV(approx. 100 PFU/well) for one hour. The infected cells were thenprepared in stock solutions, and were then two fold diluted in 2% DMEMwith 0.5% methylcellulose. 100 μl of each diluted antiviral agent isadded into each well of HIV infected cells. The treated cell cultureswere incubated in 37° C. CO₂ incubator for 24 hours. Plates ofHIV-infected cell cultures were fixed with methanol and stained with 1%crystal violet for 10 min., the dye is rinsed off with tap water. Eachplate is dried and syncytia were counted. The antiviral effect ofprotease inhibitor is calculated by 50% syncytia reduction.

Antiviral activities for the compounds of the present invention aresummarized in the following table.

    __________________________________________________________________________    Compound                           IC.sub.50                                  __________________________________________________________________________    EXAMPLE 27-30:                     >100 uM                                    Ac--Ala--Ala--(D-b-Nal)--Pip--OMe                                             EXAMPLE 28A:                       10 uM                                      DPPE--Suc--Ala--Ala--(D-b-Nal)--PiP--OMe                                      EXAMPLE 27-40:                     10 uM                                      i-Boc--(D--Phe)--(D-a-Nal)--Pip--(a-(OH)--Leu)--Val--NH.sub.2                 EXAMPLE 28E:                       2 uM                                       DPPE-Succinyl-[D--Phe]--[D-a-Nal]--Pip--[L-a-(OH)--Leu]--Val--NH.sub.2        EXAMPLE 24:                        1 uM                                       (Boc--Phe--O--CH.sub.2 --CHOH).sub.2                                          EXAMPLE 25:                        3 uM                                       (Boc--Val--Phe--O--CH.sub.2 --CHOH).sub.2                                     __________________________________________________________________________

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 35                                                 (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc-difference                                                 (B) LOCATION: 7                                                               (D) OTHER INFORMATION: carboxyl group of terminal Val-7                       residue is in amide form.                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       SerGlyAsnPheProIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: (A) peptide                                               (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc-difference                                                 (B) LOCATION: 7                                                               (D) OTHER INFORMATION: carboxyl group of terminal Val-7                       residue is in amide form                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       SerGlnAsnTyrProIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 5, 7                                                            (D) OTHER INFORMATION: Xaa=1- amino, 1-cyclohexane                            carboxylic acid (Achx); carboxyl group of terminal Val-7                      residue is in amide form.                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       SerGlnAsnTyrXaaIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 5, 7                                                            (D) OTHER INFORMATION: Xaa=1- amino, 1-cyclopropane                           carboxylic acid (Acpr); carboxyl group of terminal Val-7                      residue is in amide form.                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       SerGlnAsnTyrXaaIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 5, 7                                                            (D) OTHER INFORMATION: Xaa=1- amino, 1-cyclopentane                           carboxylic acid (Acpnt); carboxyl group of terminal Val-7                     residue is in amide form.                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       SerGlnAsnTyrXaaIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:6:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 7                                                            (D) OTHER INFORMATION: Xaa=bAla; carboxyl group of                            terminal Arg-7 residue is in amide form.                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       ThrIleLeuXaaLeuGlnArg                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 10, 13                                                          (D) OTHER INFORMATION: Xaa=5- aminoisovaleric acid(Ava);                      carboxyl group of terminal Arg-13 residue is in amide                         form.                                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       SerGlnAsnTyrProIleValThrLeuXaaThrGlnArg                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 6 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 1, 3, 4, 5, 6                                                   (D) OTHER INFORMATION: Xaa3 =phenylalanine (Phe);                             Xaa4=4- piperidine carboxylic acid (Pip); Xaa5 =                              α- hydroxyleucine(α-OH--Leu); amino group of initial Ala-1        residue is acetylated; carboxyl group of terminal Val-6                       residue is in amide form.                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       AlaAlaXaaXaaXaaVal                                                            15                                                                            (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 7                                                            (D) OTHER INFORMATION: Xaa: βalanine(bAla);                              terminal Gly-7 residue is hydroxylated.                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       AlaAlaPheXaaValValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 5, 7                                                         (D) OTHER INFORMATION: Xaa4=bAla; Xaa5=Nva                                    carboxy terminal Gly-7 residue is hydroxylated.                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      AlaAlaPheXaaXaaValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 5, 7                                                         (D) OTHER INFORMATION: Xaa4=bAla; Xaa5=α-OH-isovaleryl;                 carboxy terminal Gly-7 residue is hydroxylated.                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      AlaAlaPheXaaXaaValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 7                                                            (D) OTHER INFORMATION: Xaa: βalanine(bAla);                              carboxy terminal Gly-7 residue is a methyl ester.                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      AlaAlaPheXaaValValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 5, 7                                                         (D) OTHER INFORMATION: Xaa4=bAla; Xaa5=Nva                                    carboxy terminal Gly-7 residue is a methyl ester.                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      AlaAlaPheXaaXaaValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:14:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 5, 7                                                         (D) OTHER INFORMATION: Xaa4=bAla; Xaa5=α-OH-isovaleryl;                 carboxy terminal Gly-7 residue is a methyl ester.                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      AlaAlaPheXaaXaaValGly                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 6 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 4, 5, 6                                                         (D) OTHER INFORMATION: Xaa4=bAla; Xaa5=norvaline(Nva);                        amino terminal Ala-1 residue is modified by                                   t- butyloxycarbonyl attached to amino group; terminal                         Gly-6 residue is a methyl ester.                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                      AlaAlaPheXaaXaaGly                                                            15                                                                            (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 4                                                            (D) OTHER INFORMATION: Xaa3=L-α- naphthylalanine(L-α-Nal);        Xaa4=4- piperidine carboxylic acid (Pip); initial amino                       terminal Ala-1 residue is an acetamide; carboxy terminal                      pipecolic acid, Pip-7, is a methyl ester.                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      AlaAlaXaaXaa                                                                  (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 4                                                            (D) OTHER INFORMATION: Xaa3=L-α- naphthylalanine(L-α-Nal);        Xaa4 =4- piperidine carboxylic acid (Pip); initial amino                      terminal Ala-1 residue is an acetamide; carboxy terminal                      pipecolic acid, Pip-4, is a hydrazide.                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      AlaAlaXaaXaa                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 4                                                            (D) OTHER INFORMATION: Xaa3=L-α- naphthylalanine(L-α-Nal);        Xaa4=4- piperidine carboxylic acid (Pip); amino terminal                      Ala-1 residue is a cholic amide; carboxy terminal                             pipecolic acid, Pip-4 is a methyl ester.                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      AlaAlaXaaXaa                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 4                                                            (D) OTHER INFORMATION: Xaa3=L-α- naphthylalanine(L-α-Nal);        Xaa4=4- piperidine carboxylic acid (Pip); initial amino                       terminal Ala-1 residue is a cholic amide; carboxy                             terminal pipecolic acid, Pip-4, is a hydrazide.                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      AlaAlaXaaXaa                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 8                                                               (D) OTHER INFORMATION: carboxy terminal Val-8 residue is                      an amide.                                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      ValSerGlnAsnTyrProIleVal                                                      15                                                                            (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 4                                                            (D) OTHER INFORMATION: Xaa=4- piperidine carboxylic                           acid(Pip); Phe-2 is joined to Xaa by an isosteric bond                        having the structure --C(O)--CH2--N--; carboxy terminal                       Ile-4 is an amide.                                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      AsnPheXaaIle                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 2-3, 3, 4                                                       (D) OTHER INFORMATION: Xaa=4- piperidine carboxylic acid                      (Pip); Phe-2 is joined to Xaa by an isosteric link having                     the structure --C(O)--CH2--N--; amino terminal Asn-1 residue                  is a quinoloyl derivative; carboxy terminal Ile-4 is an                       amide.                                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      AsnPheXaaIle                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 6, 9                                                            (D) OTHER INFORMATION: Xaa=4- piperidine carboxylic acid                      (Pip); carboxy terminal Gln-9 residue is an amide.                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                      ValSerGlnAsnTyrXaaIleValGln                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 6 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 3, 6                                                            (D) OTHER INFORMATION: Xaa=decahydroisoquinoline                              carboxylic acid (Diq); Phe-2 is joined to Xaa by an                           isosteric link having the structure --CHOH--CH2--N--;                         amino terminal Asn-1 residue is a quinoloyl derivative;                       carboxy terminal Gln-6 is an amide.                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      AsnPheXaaIleValGln                                                            15                                                                            (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 6, 9                                                            (D) OTHER INFORMATION: Xaa=decahydroisoquinoline                              carboxylic acid (Diq); carboxy terminal Gln-9 residue is                      an amide.                                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      ValSerGlnAsnTyrXaaIleValGln                                                   15                                                                            (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc-difference                                                 (B) LOCATION: 1, 7                                                            (D) OTHER INFORMATION: amino terminal Ser-1 is a succinyl                     derivative; carboxyl group of terminal Val-7 residue is                       in amide form.                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      SerGlyAsnTyrProIleVal                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc-difference                                                 (B) LOCATION: 1                                                               (D) OTHER INFORMATION: amino terminal Ser-1 is a succinyl                     derivative.                                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      SerGlyAsnTyr                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 1, 3, 4                                                         (D) OTHER INFORMATION: Xaa=4- piperidine carboxylic acid                      (Pip); Phe is joined to Xaa by an isosteric link having                       the structure --CHOH--CH2--N--; amino terminal Asn-1 residue                  is a quinoloyl derivative; carboxy terminal Ile-4 is an                       amide.                                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      AsnPheXaaIle                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Misc. differences                                               (B) LOCATION: 1-2, 4                                                          (D) OTHER INFORMATION: Phe-1 is joined to Phe-2 by an                         isosteric linkage having the structure --CHOH--CH2--N--;                      carboxy terminal Ile-4 is an amide.                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      PhePheIlePhe                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Misc. differences                                               (B) LOCATION: 1-2, 4                                                          (D) OTHER INFORMATION: Phe-1 is joined to Phe-2 by an                         isosteric linkage having the structure --CHOH--CH2--N--;                      carboxy terminal Val-4 is an amide.                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      PheProIleVal                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 7, 10, 11                                                       (D) OTHER INFORMATION: Xaa7=4- NO2 phenylalanine(4-NO2--                      Phe); Xaa10=norleucine(Nle); carboxy terminal Ser-11 is                       an amide.                                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      HisLysArgAlaValLeuXaaGluAlaXaaSer                                             1510                                                                          (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Misc. differences                                               (B) LOCATION: 1, 1-2, 4                                                       (D) OTHER INFORMATION: amino terminal Phe-1 residue is a                      t- butyloxycarbonyl derivative; Phe-1 and Phe-2 are joined                    by an isosteric link having the structure --CHOH--CH2--N--;                   carboxy terminal Phe-4 is an amide.                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      PhePheIlePhe                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 1, 1-2, 4                                                       (D) OTHER INFORMATION: amino terminal Ala-1 residue is a                      t- butyloxycarbonyl derivative; Phe-1 and Pro-2 are joined                    by an isosteric link having the structure --CHOH--CH2--N--;                   carboxy terminal Val-4 is an amide.                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      PheProIleVal                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:34:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Unusual amino acids; misc. differences                          (B) LOCATION: 1, 4                                                            (D) OTHER INFORMATION: amino terminal Phe-1 residue is                        modified by succinyl attached to amino group; carboxy                         terminal Phe-4 is an amide.                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                      PhePheIlePhe                                                                  1                                                                             (2) INFORMATION FOR SEQ ID NO:35:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (v) FRAGMENT TYPE: internal                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Misc. differences                                               (B) LOCATION: 1, 4                                                            (D) OTHER INFORMATION: amino terminal Tyr-1 residue is                        modified by t-butyloxycarbonyl attached to amino group;                       carboxy terminal Gly-4 is hydroxylated.                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                      TyrProIleGly                                                                  1                                                                             __________________________________________________________________________

What is claimed is:
 1. A peptide-lipid conjugate selected from the groupconsisting of: ##STR7## wherein X represents phosphatidyl-tyrosine;phosphatidyl-serine; phosphatidyl-threonine;phosphatidyl-hydroxyproline; 1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z,wherein Y isHOOC--(CH₂)_(n) --COOH; Z is HO--(CH₂)_(n) --COOH; and n=1 to 12;Crepresents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine,1-O-acyl-sn-glycero-3-phospho-O-threonine,1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-alkyl-sn-glycero-3-diphospho-O-threonine,1-O-acyl-sn-glycero-3-diphospho-O-threonine, phosphatidyl ethanolamine,or phosphatidyl ethanolamine-W, wherein W is H₂ N--(CH₂)_(n) --COOH, andn=1 to
 12. 2. A peptide-lipid conjugate selected from the groupconsistingof:5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-Phe-C;5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-C;5(S)-X-amino-4(S)-hydroxy-6-cyclohexyl-2(R)-(phenylmethyl)hexanoyl-C;5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-Phe-C;5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(R)-(phenylmethyl)hexanoyl-Leu-C;and5(S)-X-amino-4(R)-hydroxy-6-cyclohexyl-2(R)-(phenylmethyl)hexanoyl-C;whereinX represents phosphatidyl-tyrosine; phosphatidyl-serine;phosphatidyl-threonine; phosphatidyl-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z, wherein Y isHOOC--(CH₂)_(n) --COOH and Z is HO--(CH₂)_(n) --COOH; and n=1 to 12; andC represents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine,1-O-acyl-sn-glycero-3-phospho-O-threonine,1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-alkyl-sn-glycero-3-diphospho-O-threonine,1-O-acyl-sn-glycero-3-diphospho-O-threonine, phosphatidyl ethanolamine,or phosphatidyl ethanolamine-W, wherein W is H₂ N--(CH₂)_(n) --COOH, andn=1 to
 12. 3. A peptide-lipid conjugate selected from the groupconsistingof:5(S)-X-Amino-4(S)-hydroxy-6-cyclcohexyl-2(R)-isopropyl-hexanoyl-Ile-C;5(S)-X-Amino-4(R)-hydroxy-6-cyclcohexyl-2(S)-isopropyl-hexanoyl-Ile-C;5(S)-X-Amino-4(S)-hydroxy-6-phenyl-2(R)-isopropyl-hexanoyl-Ile-C; and5(S)-X-Amino-4(R)-hydroxy-6-phenyl-2(S)-isopropyl-hexanoyl-Ile-C;whereinX represents phosphatidyl-tyrosine; phosphatidyl-serine;phosphatidyl-threonine; phosphatidyl-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z, wherein Y isHOOC--(CH₂)_(n) --COOH and Z is HO--(CH₂)_(n) --COOH; and n=1 to 12; andC represents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine,1-O-acyl-sn-glycero-3-phospho-O-threonine,1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-alkyl-sn-glycero-3-diphospho-O-threonine,1-O-acyl-sn-glycero-3-diphospho-O-threonine, phosphatidyl ethanolamine,or phosphatidyl ethanolamine-W, wherein W is H₂ N--(CH₂)_(n) --COOH, andn=1 to
 12. 4. A peptide-lipid conjugate selected from the groupconsistingof:(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-Pro-C;(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-Pro-C;(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-C;(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-Ile-C;(3(S)-X-Leucyl-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-Pro-Ile-C;(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-N-1,2,3,4-tetrahydro(R,S)isoquinolinecarboxyl-C;(3(S)-X-Asparaginyl)-amino-2(R,S)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C;(3(S)-X-Asparaginyl)-amino-2(R)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C; and(3(S)-X-Asparaginyl)-amino-2(S)-hydroxy-4-phenylbutyl-N-decahydro-3(S)-isoquinolinecarboxyl-C;wherein X represents phosphatidyl-tyrosine;phosphatidyl-serine; phosphatidyl-threonine;phosphatidyl-hydroxyproline; 1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z, wherein Y isHOOC--(CH₂)_(n) --COOH and Z is HO--(CH₂)_(n) --COOH; and n=1 to 12; andC represents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine,1-O-acyl-sn-glycero-3-phospho-O-threonine,1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-alkyl-sn-glycero-3-diphospho-O-threonine,1-O-acyl-sn-glycero-3-diphospho-O-threonine, phosphatidyl ethanolamine,or phosphatidyl ethanolamine-W, wherein W is H₂ N--(CH₂)_(n) --COOH, andn=1 to
 12. 5. A peptide-lipid conjugate selected from the groupconsistingof:4(S)-(X-Alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C;4(S)-(X-Alanyl-alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C;4(S)-(X-Seryl-alanyl-alanyl)amino-3(S)-hydroxy-5-phenyl-pentanoyl-Val-Val-C4(S)-(X-Alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C;4(S)-(X-Alanyl-alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C;and4(S)-(X-Seryl-alanyl-alanyl)amino-3(R)-hydroxy-5-phenyl-pentanoyl-Val-Val-C;whereinX represents phosphatidyl-tyrosine; phosphatidyl-serine;phosphatidyl-threonine; phosphatidyl-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z, wherein Y isHOOC--(CH₂)_(n) --COOH and Z is HO--(CH₂)_(n) --COOH; and n=1 to 12; andC represents H, OH, OMe, NH₂, NH--R where R represents C₁₋₁₂ alkyl,benzyl, substituted benzyl, (CH₂)_(n) -phenyl where n=1 to 12, 2-methylpyridyl, phosphatidyl-tyrosine, phosphatidyl-serine,phosphatidyl-threonine, phosphatidyl-hydroxyproline,1-O-alkyl-sn-glycero-3-phospho-O-tyrosine,1-O-acyl-sn-glycero-3-phospho-O-tyrosine,1-O-alkyl-sn-glycero-3-phospho-O-serine,1-O-acyl-sn-glycero-3-phospho-O-serine,1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine,1-O-acyl-sn-glycero-3-phospho-O-threonine,1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine,1-O-acyl-sn-glycero-3-diphospho-O-tyrosine,1-O-alkyl-sn-glycero-3-diphospho-O-serine,1-O-acyl-sn-glycero-3-diphospho-O-serine,1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline,1-O-alkyl-sn-glycero-3-diphospho-O-threonine,1-O-acyl-sn-glycero-3-diphospho-O-threonine, phosphatidyl ethanolamine,or phosphatidyl ethanolamine-W, wherein W is H₂ N--(CH₂)_(n) --COOH, andn=1 to
 12. 6. A peptide-lipid conjugate according to claim 1 selectedfrom the group consisting of:i-Boc-Tyr-Pro-Ile-Gly-DPPE;DPPA-Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH₂ ;DPPA-Ser-Gln-Asn-Tyr-Acpnt-Ile-Val-NH₂ ;DPPE-Succinyl-Ala-Ala-(D-b-Nal)-Pip-OMe;DPPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂ ;DMPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Pip-Ile-Val-Gln-NH₂ ;DMPE-Succinyl-Val-Ser-Gln-Asn-Tyr-Diq-Ile-Val-Gln-NH₂ ;DPPE-Succinyl-(D-Phe)-(D-a-Nal)-Pip-(L-a-(OH)-Leu)-Val-NH₂ ;DPPE-Succinyl-Phe(CHOH--CH2N)Phe-Ile-Phe-NH₂ ; ##STR8## X-Phe(CHOH--CH₂N)Phe-Ile-Phe-NH₂ ; and X-Phe(CHOH--CH₂ N)Pro-Ile-Val-NH₂,wherein DMPEis dimyristoyl phosphatidyl ethanolamine; DPPE is dipalmitoylethanolamine; and DPPA is dipalmitoyl phosphatidic acid; and Ser(DPP) isdipalmitoyl phosphatidyl serine.
 7. A peptide lipid conjugate of theformula

    iBoc-Phe-Pip-Val-DPPE.


8. 8. A peptide-lipid conjugate selected from the group consistingof:X-Val-Phe-Nva-(cyclohexylmethyl(4,4,5,5-tetramethyl-1,3,2-dioxoborlan-2-yl)methylamide;X-Val-Phe-Nva-(cyclohexylmethyl, dihydroxyboronyl)methylamide;X-Val-Phe(L-a-Nal)-Nva-(cyclohexylmethyl(4,4,5,5-tetramethyl-1,3,2-dioxoborlan-2-yl)methylamide;X-Val-Phe(L-a-Nal)-Nva-(cyclohexylmethyl, dihydroxyboronyl)methylamide;X-Nva(cyclohexylmethyl, dihydroxyboronyl)methylamide; andX-Val(cyclohexylmethyl, dihydroxyboronyl)methylamide;wherein: Xrepresents phosphatidyl-tyrosine; phosphatidyl-serine;phosphatidyl-threonine; phosphatidyl-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-tyrosine;1-O-acyl-sn-glycero-3-phospho-O-tyrosine;1-O-alkyl-sn-glycero-3-phospho-O-serine;1-O-acyl-sn-glycero-3-phospho-O-serine;1-O-alkyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-phospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-phospho-O-threonine;1-O-acyl-sn-glycero-3-phospho-O-threonine;1-O-alkyl-sn-glycero-3-diphospho-O-tyrosine;1-O-acyl-sn-glycero-3-diphospho-O-tyrosine;1-O-alkyl-sn-glycero-3-diphospho-O-serine;1-O-acyl-sn-glycero-3-diphospho-O-serine;1-O-alkyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-acyl-sn-glycero-3-diphospho-O-hydroxyproline;1-O-alkyl-sn-glycero-3-diphospho-O-threonine;1-O-acyl-sn-glycero-3-diphospho-O-threonine; phosphatidylethanolamine-Y; phosphatidic acid, or phosphatidic acid-Z,wherein Y isHOOC--(CH₂)_(n) --COOH; Z is HO--(CH₂)_(n) --COOH; and n=1 to 12.